渐进性梯度孔径骨软骨支架的制备及细胞相容性研究  

Preparation of progressive gradient-aperture osteochondral scaffold and its cytocompatibility evaluation

作　　者：肖红利 邓江 韩子冀[1] 黄文良 熊坤 张勇 Xiao Hongli;Deng Jiang;Han Ziji;Huang Wenliang;Xiong Kun;Zhang Yong(Department of Orthopedics,the First People's Hospital of Guiyang,Guiyang 550000,China;Department of Orthopedics,the Third Affiliated Hospital of Zunyi Medical University(the First People's Hospital of Zunyi),Zunyi 563000,China)

机构地区：[1]贵阳市第一人民医院骨科,550000 [2]遵义医科大学第三附属医院(遵义市第一人民医院)骨科,563000

出　　处：《中华老年医学杂志》2020年第4期456-461,共6页Chinese Journal of Geriatrics

基　　金：国家自然科学基金委员会资助项目(81660367);贵州省科学技术基金项目(黔科合基础(2016)1420号)。

摘　　要：目的:制备一种用于骨软骨修复的丝素蛋白(SF)-壳聚糖(CS)-纳米羟基磷灰石(nHAp)渐进性梯度孔径支架。方法:将SF溶液、CS溶液及nHA悬液等比例混合,采用离心、真空冷冻干燥及化学交联法,3次塑形,制备成渐进性梯度孔径骨软骨(OC)支架-1(2%)、支架-2(3%)、支架-3(4%)。检测各组支架一般情况、孔隙率、热水溶失率、吸水膨胀率、压缩吸水膨胀率、溶失后吸水膨胀率、力学性能、支架内部结构观察及孔径大小。复苏培养大鼠骨髓间充质干细胞(BMSCs),制备支架浸提液,CCK-8法检测浸提液对BMSCs增殖活性的影响。将BMSCs与支架共同培养,观察支架周围细胞的分布及形态。结果:各组支架形态规则,孔隙率均大于80%;随材料浓度增高,支架吸水膨胀率逐渐减小(P<0.05),压缩后支架-1吸水膨胀率明显降低(P<0.05);各组支架热水溶失率无明显差异(P>0.05),支架在体外完全溶失分别需要65.9、60.9、73.9周;三组支架弹性模量分别为(0.0955、0.1762、0.3468)MPa;扫描电子显微镜(SEM)显示各组支架内部均呈蜂窝状,孔隙形状规则、互通,从支架成软骨侧到成骨侧孔隙分布逐渐密集、孔隙直径逐渐减小(P<0.05),nHAp含量亦逐渐增多。各组支架浸提液对BMSCs的生长、增殖均无明显毒性。BMSCs与支架共同培养第5 d时,细胞生长状态良好,未出现明显的细胞死亡或形态异常。结论:本研究成功制备出渐进性梯度孔径OC支架,支架各项物理性能及生物相容性良好,有望成为修复OC缺损的新型仿生复合支架材料。Objective To prepare a progressive gradient-aperture scaffold composed of silk fibroin(SF)-chitosan(CS)-nano-hydroxyapatite(nHAp)for osteochondral repair.Method The SF solution,CS solution and nHA suspension were mixed in vitro at equal proportions.The progressive gradient osteochondral(OC)scaffold-1(2%),scaffold-2(3%)and scaffold-3(4%)was respectively prepared by using centrifugation,vacuum freeze-drying,chemical cross-linking and three shaping steps.General conditions,porosity,hot water dissolution rate,water swelling rate,compression water swelling rate,water swelling rate after dissolution,mechanical properties,internal structure observation and pore size were measured.Rat bone marrow mesenchymal stem cells(BMSCs)were cultured and the scaffold extract was prepared.The effect of scaffold extract on the proliferation of BMSCs was detected by the cell counting kit-8(CCK-8)method.BMSCs were co-cultured with the scaffold,and the distribution and morphology of the cells around the scaffold were observed.Results The structure of scaffold was regular in each group and the porosity was more than 80%.Along with the increase of the material concentration,the water swelling rate of the scaffold was decreased gradually(P<0.05).Compared with before compression,the water swelling rate of scaffold-1 was decreased after compression(P<0.05).There was no significant difference in the hot water dissolution rate among all groups(all P>0.05),and the complete dissolution of the scaffold-1,scaffold-2 and scaffold-3 in vitro required 65.9,60.9,and 73.9weeks,respectively.The elastic modulus of scaffolds in above three groups were 0.0955,0.1762 and 0.3468 MPa,respectively.The examination results of scanning electron microscope(SEM)showed that the internal structure of scaffold was honeycomb in each group,the pore shape was regular,which showed an inter-connected pore network.The pore distribution was gradually dense and the pore diameter gradually decreased from the cartilage side to the osteogenic side(P<0.05),and the nHAp content inc

关 键 词：软骨 组织支架 

分 类 号：R6[医药卫生—外科学] R3[医药卫生—临床医学]